Arylhydrazides



United States Patent 3,274,232 ARYLHYDRAZIDES Jack W. Hinman, Portage, and Ronald B. Kelly, Cooper Township, Kalamazoo County, Mich., assignors to The Upjohn Company, Kalamazoo, Mich., a corporation of Michigan No Drawing. Original application Aug. 4, 1961, Ser. No. 129,222. Divided and this application Dec. 23, 1964, Ser. No. 420,786

4 Claims. (Cl. 260-471) This application is a division of application Serial No. 129,222, filed August 4, 1961, now abandoned.

The present invention is directed to novel arylhydrazide intermediates utilized in the preparation of novel aamino dibasic acid hydrazides.

The novel a-amino dibasic acid hydrazides can be represented by the formula:

wherein R represents hydrogen, carboxy, alkyl of 1 to 4 carbon atoms, inclusive, e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and the like, halogen, e.g., bromo, chloro, fluoro, and iodo, and alkoxy of 1 to 4 carbon atoms, inclusive, e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, and the like; R represents hydrogen and alkyl of 1 to 4 carbon atoms, inclusive, e.g., methyl, ethyl, propyl, butyl, isopropyl, isobutyl, and the like, and n is an integer from zero to 7, inclusive. The structural formula has been written in zwitterion form, as is commonly done in the case of u-amino acids.

As is evident from Formula I ,the compounds of the present invention can exist in different stereoisomeric configurations due to the presence of the asymmetric carbon atom in the 2-position, i.e., the carbon atom to which the amino group is attached. It is, therefore, to be understood that the L-, the D-, and the DL-forms are within the scope of the present invention.

The a-amino dibasic acid hydrazides prepared by the novel process of the present invention are potent inhibitors of S-hydroxytryptophan decarboxylase, the enzyme system which is responsible for the conversion of S-hydroxytryptophan to serotonin in the body [Udenfriend et al., J. Biol. Chem. 224, 803 (1957)]. S-hydroxytryptophan is known to be the precursor of serotonin released in the brain [serotonin itself does not cross the bloodbrain barrier; see Fed. Proc. 15, 402 and 493 (1957)]. While the precise role which serotonin plays in the complex series of chemical reactions occurring in the brain, both normal and abnormal, is not yet clear, it is becoming increasingly evident that serotonin does have a significant effect on the manner in which the brain functions. The a-amino dibasic acid hydrazides of the present invention thus provide a regulator for the supply of serotonin to the brain and can thus be utilized in reducing hyperactivity in animals and making them more suitable for handling and transportation purposes.

The aforesaid inhibitory activity of a-amino dibasic acid hydrazides of the present invention also makes them valuable as laboratory agents. Thus, they can be used in the research laboratory to inhibit selectively S-hy- 3 ,Z74,Z3Z Patented Sept. 20, 1966 droxytryptophan decarboxylase in mixed enzyme systems. They can also be used in the characterization and quantitative determination of S-hydroxytryptophan decarboxylase in the laboratory.

L-glutamic acid 5-[2-(a-hydroxy-p-tolyl)hydrazide], an a-amino dibasic acid hydrazide, has been disclosed by Levenberg [J. Amer. Chem. Soc. 83, 503, (1961)] who proposed the trivial name agaritine, which will be used hereinafter. Agaritine has the following structural formula:

wherein R and n have the values represented above and R represents an amino protective group, e.g., carbowith a phenylhydrazine having the formula:

(III) carboallyloxy and tosyl wherein R represents hydrogen, methylol, carboxy, alkyl of 1 to 4 carbon atoms, inclusive, e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and the like; halogen, e.g., bromo, chloro, fluoro, and iodo, and alkoxy of 1 to 4 carbon atoms, inclusive, e.g., methoxy, ethoxy, propoxy, isopropoxy, butoXy, isobutoxy, and the like, to produce a novel arylhydrazide intermediate having the formula:

II I @NH-NH-C-(ChHzQ-(f-C 0 on 1 in (IV) R:

wherein R R R and n have the values represented above, and then removing the amino protective group (R from the novel arylhydrazide intermediate (Formula IV), to produce an u-amino dibasic acid hydrazide having the formula:

hydrochloride with benzyl chloroformate to produce 5- ethyl L-N carbobenzyloxyglutamate, reacting the latter compound with hydrazine hydrate to produce L-N-carbobenzyloxyglutamic acid S-hydrazide, which when reacted with nitrous acid (generated in situ with sodium nitrite and hydrochloric acid) results in the preparation of L-N- carbobenzyloxyglutamic acid S-azide.

- By susbtituting other w-alkyl esters of a-arnino dibasic acids (i.e., monoalkyl esters in which the carboxyl group farthest removed from the amino group is esterified) in the process of US. Patent No. 2,723,973, such as the w methyl and w-ethyl esters of aminomalonic acid, aspartic acid, Z-methylaspartic acid, 3,3-dimethylaspartic acid, 2- propylaspartic acid, 3-butylaspartic acid, 3-methylaspartic acid, 2-methylglutamic acid, 3-methylglutamic acid, 4- methylglutamic acid, Z-arninohexanedioic acid, 2-aminoheptanedioic acid, 2-amino-octanedioic acid, 2-aminononanedioic acid, and Z-aminodecanedioic acid for S-ethyl L-glutamate, the following hydrazides and azides, respectively, are prepared:

N-carbobenzyloxyaminomalonic acid hydrazide, N-

carbobenzyloxyaminomalonic acid azide;

N-carbobenzyloxyaspartic acid 4-hydride, N-carbobenzyloxyaspartic acid 4-azide;

N-carbobenzyloxy-Z-methylaspartic acid 4-hydrazide, N-carbobenzy1oXy-2-methylaspartic acid 4-azide;

N-carbobenzyloXy-3,3-dimethylaspartic acid 4-hydrazide, N-carbobenzyloXy-3,3-dirnethylaspartic acid 4-azide;

N-carbobenzyloxy-Z-propylaspartic acid 4-hydrazide, N-carbobenZyloXy-2-propylaspartic acid 4-azide; N-carbobenzyl'oXy-3-butylaspartic acid 4-hydrazide, N-

carbobenzyloxy-3-butylaspartic acid 4-azide;

N-carbobenzyl-oxy-3-methylaspartic acid 4-hydrazide, N-carbobenzyloXy-3-methylaspartic acid 4-azide;

N-carbobenzyloxy-Z-methylglutamic acid S-hydrazide, N-carbobenzyloXy-2-methylglutamic acid 5-azide;

N-carbobenzyloxy-3-methylglutamic acid S-hydrazide, N-carbobenzyloXy-3-methy1glutamic acid 5-azide;

N-carbobenzyloXy-4-methylglutarnic acid S-hydrazide, N-carbobenzyloxy-4-methylglutamic acid S-azide;

N-carbobenzyloxy-Z-aminohexanedioic acid 6- hydrazide, N-carbobenzyloxy-Z-arninohexanedioic acid 6-azide;

N-carbobenzyloxy-Z-aminoheptanedioic acid 7-hydrazide, N-oarbobenzyloxy-Z-aminoheptanedioic acid 7-azide;

N-carbobenzyloxy-2-aminooctanedioic acid 8-hydrazide, N-carbobenzyloxy-Z-aminooctanedioic acid 8-azide;

N-carbobenzyloxy-Z-amirrononanedioic acid 9-hydrazide, N-carbobenzyloxy-Z-aminononanedioic acid 9-azide;

N-carbobenzyloxy-Z-aminodecanedioic acid IO-hydrazide, N-carbobenzyloxy-Z-aminodecanedioic acid 10 azide.

By employing the foregoing w-alkyl esters of m-arnino dibasic acids in the process of U8. Patent No. 2,723,973, but substituting ally-l chlorformate and p-toluenesulfonyl chloride (tosyl chloride) for benzyl chloroformate, the following hydrazides and azides are prepared;

N-carboallyloxyglutamic acid S-hydrazide, N-tosylglutamic acid S-hydrazide,-N-carboallyloxyglutamic acid S-azide, N-tosylglutarnic acid 5-azide;

N-carboallyloxyaminomalonic acid hydrazide, N-tosylaminomalronic acid hydrazide, N-carboallyloxyaminomalonic acid azide, N-tosylaminomalonic acid azide;

N-canboallyloxyaspartic acid 4-hydrazide, N-tosylaspartic acid 4-hydrazide, N-carboallyloxyasp-artic acid 4-azide, N-tosylaspartic acid 4-azide;

N-carboallyloxy-Z-methylaspartic acid 4-hydrazide, N- tosy1-2-methylaspa1tic acid 4-hydrazide, N-carboallyloxy-2-methylaspartic acid 4-azide, N-tosy1-2- methylaspartic acid 4-azide;

N-carboa1lyl oXy-3,3-dimethylaspartic acid 4-hydrazide,

N-tosyl-3,3-dimethylaspartic acid 4-hydrazide N- carboa1lyloxy-3,3-dimethylaspartic acid 4-azide, N- tosyl-3,3-dimethylaspartic acid 4-azide;

N-carboallyloxy-Z-propylaspartic acid 4-hydrazide, N-

tosyl-2-propylasparatic acid-4 hydrazide, N-carboallyloxy-2-propylasparatic acid 4-azide, N-tosyl-Z-propylaspartic acid 4-azide;

N-carboallyloxy-3-butylaspartic acid 4-hydrazide, N-

tosyl-3-butylaspartic acid 4-hydrazide, N-carboallyloxy- 3-butylaspartic acid 4-azide, N-tosyl-3-butylaspartic acid 4-azide;

N-carboallyloxy-3-nrethylaspartic acid 4-hydrazide, N-

tosyl-3-methylasparatic acid 4-hydrazide, N-carboallyloxy-3-methylasparatic acid 4-azide, N-tosy1-3-methylaspartic acid 4-azide;

N-carboallyloxy-2-methylglutamic acid S-hydrazide, N-

t-osyl-Z-methylglutamic acid S-hydrazide, N-carboallyloxy-2-methylglutamic acid S-azide, N-tosyl-Z-methylglutamic acid S-azide;

N-carboallyloXy-3-methylglutamic acid S-hydrazide, N-

tosyl-3-methylglutamic acid S-hydrazide, N-carboallyloxy-3-methylglutamic acid S-azide, N-tosyl-3-methylglutamic acid S-aZide;

N-carboallyloxy-4-methylglutamic acid S-hydrazide, N-

tosyl-4-methylglutamic acid S-hydrazide, N-carboallyloxy-4-methylglutamic acid S-azide, N-tosyl-4-methylglutamic acid S-azide;

N-carboallyloxy-2-aminohexanedioic acid 6-hydrazide,

N-tosyl-2-aminohexanedioic acid 6-hydrazide, N-carboallyloxy-Z-aminohexanedioic acid 6-azide, N-tosyl- Z-aminohexanedioic acid 6-azide;

N-carboallyloxy-Z-aminoheptanedioic acid 7-hydrazide,

N-tosyl-Z-aminoheptanedioic acid 7-hydrazide, N-carboallyloxy-Z-aminoheptanedioic acid 7-azide, N-tosyl- 2-aminoheptanedioic acid 7-azid'e;

N-carboallyloXy-Z-aminooctanedioic acid S-hydrazide,

N-tosyl-2-aminooctanedioic acid 8-hydrazide, N-ca-r' boallyloxy-Z-aminooctanedioic acid 8-azide, N-tosyl- 2-aminooctane-dioic acid 8-azide;

N-carboallyloxy-2-aminononanedioic acid 9-hydrazide, N-tosyl-2-aminononanedioic acid 9-hydrazide, N carboallyloxy-Z-aminononanedioic acid 9-azide, N-tosyl- Z-aminononanedioic acid 9-azide;

N-carboallyloxy-Z-aminodecanedioic acid IO-hydrazide,

N-tosyl-2-aminodecanedioic acid IO-hydrazide, N-carboallyloxy-Z-aminodecanedioic acid 10-azide, and

N-tosyl-2-aminodecanedioic acid 10-azide.

Generally speaking, the azides (Formula II) are not particularly stable; Therefore, it is preferred not to isolate them, but to prepare solutions of them (e.g., ether or chloroform solutions), which are then used in further synthesis as soon thereafter as is practicable.

In carrying out the process of the present invention, the azide (Formula II) is reacted with the phenylhydrazine (Formula III) in the presence of an inert solvent, e.g., ether, ethyl acetate, chloroform, methylene chloride, and the like. The reaction is advantageously carried out at a temperature between about 15 C. and about 10 C., preferably between about -5 C. and about C. Upon completion of the reaction, the resulting novel arylhydrazide intermediate (Formula IV) can be isolated and purified if so desired, using conventional techniques, such as filtration, solvent extraction, evaporation, recrystallization, and the like.

The novel arylhydrazide intermediate is then subjected to hydrolysis or hydrogenolysis to remove the protective group (R in Formula IV) and thus obtain the desired OC-flfl'lll'lO dibasic acid hydrazide (Formula V).

Preferably, the carbobenzyloxy protective group is removed by catalytic hydrogenolysis utilizing a palladium catalyst, such as palladium-barium sulfate, palladiumcharcoal, palladium black, and the like, with a 10% palladium-charcoal catalyst being preferred. Other concentrations between about 1% and 50% can also be em ployed, with a somewhat shorter reaction time being required at the higher concentration. Inert solvents in which the hydrogenolysis can be carried out include, among others, lower alkanols, such as aqueous or anhydrous methanol, ethanol, and isopropanol. After the hydrogenolysis has been completed, the resulting a-amino dibasic acid hydrazide (Formula V) can be isolated and purified by means of conventional techniques, such as filtration to remove the catalyst, chromatography, removal of solvent, recrystallization, and the like. Sodium and liquid ammonia, phosphonium iodide, and hydrogen halide in glacial acetic acid can also be employed to remove the carbobenzyloxy group as disclosed in U.S. Patent 2,723,973.

Preferably, the carboallyloxy protective group is removed by catalytic hydrogenolysis using platinum oxide catalyst in acidified ethanol as disclosed by Milne et al., J. Amer. Chem. Soc. 79, 637-39, 1957. This process can also be employed to remove the carbobenzyloxy protective group.

Preferably, the tosyl protective group is removed with sodium and liquid ammonia as disclosed by Milne et al., J. Amer. Chem. Soc. 79, 639-44, 1957. Alternatively, it can be removed by other procedures disclosed by the same authors, e.g., by means of Raney nickel, or hydriodic acid in glacial acetic acid, or hydrobromic acid and phenol.

The following examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

EXAMPLE l.L-GLUTAMIC ACID 5 Z-PHENYLHYDRAZIDE) A. L-N-carbobenzyloxyglutamic acid S-hydrazide To a stirred solution of 53.4 g. of S-methyl L-glutamate, 59 g. of sodium bicarbonate, and 720 ml. of water, cooled to 05 C. in an ice bath, was added 60 ml. of benzyl chloroformate over a period of 25 minutes. Stirring and cooling was continued for 1.5 hours, the cooling bath was removed, and the mixture was stirred for an addiitonal 2 hours. The mixture was extracted with three 700-ml. portions of ether, acidified with concentrated hydrochloric acid, and extracted with three 1000-ml. portions of ethyl acetate. The extracts were washed with 250 m1. of water containing 1.5 g. of sodium bicarbonate, combined, and evaporated to a syrup under reduced pressure. The syrup was crystallized twice from carbon tetrachloride to produce 40 g. of S-methyl L-N-carbobenzyloxyglutamate melting at 70 C.

17.0 g. of S-methyl L-N-carbobenzyloxyglutamate was dissolved in 40 ml. of absolute ethanol and the solution was treated, with cooling, with 18 ml. of hydrazine hydrate (99100%). The reaction mixture was left in a stoppered flask at 25 C. for 48 hours. It was then concentrated under reduced pressure at 38 C. to 15 ml. Water 130 ml.) was added to the residue and the aqueous solution was cooled in an ice bath while being treated with concentrated hydrochloric acid. When the solution became acidic, the product crystallized, was collected on a funnel and washed with water, methanol and ether. The dried product, L-N-carbobenzyloxyglutamic acid 5- hydnazide, weighed 13.0 g. and melted at 179180 C.

Analysis.--Calcd. for C H N O C, 52.97; H, 5.80; N, 14.23. Found: C, 52.71; H, 5.62; N, 14.72.

B. L-N-carbobenzyloxyglutamic acid 5-azide A solution of 54.4 g. (0.18 mole) of L-N-carbobenzyloxyglutamic acid S-hydrazide in 300 ml. of 3 N hydrochloric acid and ml. of water was cooled in an ice bath. The cooled solution was covered with cold (10 C.) ether and the mixture was stirred while a cold solution of 15.2 g. (0.22 mole) of sodium nitrite in 100 ml. of water was added over a period of 15 min. The lower layer was separated and extracted with two 1.0-]. portions of cold (10 C.) ether. The ether extracts were washed with two 500-ml. portions of ice water, combined, and dried briefly over anhydrous sodium sulfate. There was thus obtained a cold (0 C.) ether solution of L-N-carbobenzyloxyglutamic acid 5-azide.

C. L-N-carbobenzyloxyglutamic acid 5 Z-phenylhydmzide) The above ether solution of L-N-carbobenzyloxyglutamic acid 5-azide was added portionwise with stirring over a period of 20 minutes to a solution of 43.2 g. (0.40 mole) of phenylhydrazine in 500 ml. of ether cooled in an ice bath. The reaction mixture was cooled in the ice bath for 8 hours and then kept at 25 C. for 8 hours. The resulting precipitate (76.2 g.) was collected and washed with ether and was then dissolved in a mixture of 1000 ml. of ethyl acetate and 500 ml. of l N sulfuric acid. The layers were separated and the aqueous layer was extracted with two 1000-ml. portions of ethyl acetate. The ethyl acetate extracts were washed with two 500-ml. portions of l N sulfuric acid, then with two 250-ml. portions of Water. The original ethyl acetate layer and the ethyl acetate extracts were dried over anhydrous sodium sulfate, combined, and evaporated under reduced pressure at 38 C. to a thick syrup which was crystallized from 60% methanol. The yield of crystalline material, L-N- carbobenzyloxyglutamic acid S-(Z-phenylhydrazide), was 47.3 g. (70%). The product, after being recrystallized from ethanol-water (65%), melted at 7080 C., slowly resolidified between 100-140 C., and finally melted at 148149 C.

Analysis.--Calcd. for C H N O C, 61.44; H, 5.70; N, 11.32. Found: C, 61.12; H, 5.86; N, 11.25.

D. L-glutamic acid S-(Z-phenylhydrazide) A solution of 5.0 g. of L-N-carbobenzyloxyglutamic acid S-(Z-phenylhydrazide) in 200 ml. of 50% methanol was hydrogenolyzed in the presence of 500 mg. of palladium-charcoal catalyst (10%) for one hour at 18 p.s.i. (gauge). The catalyst was removed by filtration and washed well with warm 50% methanol and warm water. The combined washings and filtrate (1.0 l.) were evaporated under reduced pressure at 36 C. to a volume of 300 ml., then extracted with 200-ml. portions of methylene chloride, ethyl acetate, and ether. The extracted aqueous solution was concentrated under reduced pressure at 38 C. to a volume of 100 ml. Ethanol (100 ml.) was added to the concentrate and 1.99 g. (57%) of crystalline product, L-glutamic acid 5-(2-phenylhydrazide), was obtained. An analytical sample prepared by two recrystallizations from 50% ethanol melted at 203 C.; [a] -|24 (c.=0.41 in 0.5 N hydrochloric acid).

Analysis.Calcd. for C H N O C, 55.68; H, 6.37; N, 17.71. Found: C, 55.50; H, 6.16; N, 17.73.

Following the procedure of Example 1, but starting with S-methyl D-glutamate instead of S-methyl L-glutamate, the following compounds are obtained: S-methyl D-N-carbobenzyloxyglutamate and D-N-carbobenzyloxyglutamic acid -hydrazide in Part A; D-N-carbobenzyloxyglutamic acid 5-azide (ether solution) in Part B; D-N- carbobenzyloxyglutarnic acid 5-(2-phenylhydrazide) in Part C; and D-glutamic acid 5-(2-phenylhydrazide) in Part D.

In like manner, but starting with S-methyl D,L-glutamate, the corresponding derivatives of D,L-glutamic acid are obtained.

EXAMPLE 2.L-GLUTAMIC ACID 5-(2-p- TOLYLHYD-RAZIDE) A. L-N-carbobenzyloxyglutamic acid 5-(2-p-tolylhydrazide) An ether solution of L-N-carbobenzyloxyglutamic acid 5-azide was prepared from 21.5 g. of L-N-carbobenzyloxyglutamic acid S-hydr-azide in the manner described in Example 1, and stored in an acetone-solid carbon dioxide bath. The solution was added, over a period of one hour, to a stirred solution of 18.3 g. (0.15 mole) of p-tolylhydrazine in 250 ml. of ether. During the addition of the azide solution, the p-tolylhydrazine solution was cooled in an ice bath. The ice bath was removed and the reaction mixture was kept at 25 C. for 22 hours. The precipitate which formed was collected, washed well with ether, and dissolved in a mixture of 1.0 l. of methylene chloride and 500 ml. of 1 N hydrochloric acid. The aqueous layer was separate-d and extracted with four 1.0-l. portions of methylene chloride. The methylene chloride extracts were washed with 500 ml. of 1 N hydrochloric acid and with two 500-rnl. portions of water. The combined methylene chloride extracts and original methylene chloride layer were evaporated under reduced pressure at 36 C. The residue was crystallized from 75% methanol to yield 15.0 g. of L-N-carbobenzyloxyglutamic acid 5-(2-p-tolylhydrazide). Upon recrystallization from methanol-ether, then from 80% ethanol, an analytical sample which was solvated with ethanol had a melting point of 153155 C.

Analysis.Calcd. for C20H23N305'CH3CH2OHZ C, 61.24; H, 6.77; N, 9.74. Found: C, 61.16; H, 6.18; N, 10.01.

In the same manner as shown in Example 2, Part A,

L-N-carbobenzyloxyglutamic acid 5-(2-o-ethylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5-(2-m-propylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5-(2-p-butylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5- (2-o-isopropylphenylhydrazide), and

L-N-carbobenzyloxyglutamic acid 5-(2-m-isobutylphenylhydrazide are prepared by using o-ethylphenylhydrazine,

m-propylphenylhydrazine, p-butylphenylhydrazine, o-isopropylphenylhydrazine, and m-isobutylphenylhydrazine,

respectively, instead of p-tolylhydrazine.

B. L-glutamic acid 5-(2-p-tolylhydrazide) A solution of 3.85 g. (0.01 mole) of L-N-carbobenzyloxyglutamic acid 5-(2-p-tolylhydrazide) in 250 ml. of 60% ethanol was hydrogenolyzed in the presence of 250 mg. of palladium-charcoal catalyst for one hour at 17 p.s.i. (gauge). The catalyst was removed by filtration and washed with warm water. The combined filtrate and washings were evaporated under reduced pressure at 38 C. to a volume of 100 m1. Water (150 ml.) was added to dissolve the resulting precipitate and the aqueous solution was extracted with two 150-ml. portions of ethyl acetate and then with 150 ml. of ether. The organic extracts were washed with two 100-ml. portions of water. The extracted aqueous solution and washings were combined and evaporated under reduced pressure at 38 C. to a volume of 200 ml. Ethanol (200 ml.) was added and the resulting solution deposited 1.40 g. of product when cooled in the refrigerator. An additional 0.54 g. of product was obtained from the mother liquors by concentration to ml. and addition of 30 ml. of ethanol and ml. of n-butyl alcohol. The yield of crystalline product, L-glutamic acid 5-(2-p-tolylhydrazide), was 77% and the product melted at l83184 C.; +25 (c.=0.88 in 0.5 N hydrochloric acid).

Analysis.Calcd. for C H N O C, 57.29; H, 6.82; N, 16.71; 0, 19.18. Found: C, 57.42; H, 6.69; N, 16.25; 0, 19.34.

In the same manner as shown in Example 2, Part B,

L-glutarnic acid 5-(2-o-ethylphenylhydrazide), L-glutamic acid 5-(2-m-propylphenylhydrazide), L-glutamic acid 5-(2-p-butylphenylhydrazide), L-glutamic acid 5-(2-o-isopropylphenylhydrazide), and L-glutamic acid 5-(2-m-isobutylphenylhydrazide) are prepared by using.

L-N-carbo'benzyloxyglutamic acid 5-(2-o-ethylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5-(2-m-propylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5-(2-p-butylphenylhydrazide),

L-N-carbobenzyloxyglutamic acid 5-(2-o-isopropylp'henylhydrazide), and

L-N-carbobenzyloxyglutamic acid 5-(2-m-isobutylpheny1- hydrazide), respectively, instead of L-N-carbobenzyloxyglutamic acid 5-(2-p-tolylhydrazide).

EXAMPLE 3.--L-GLUTAMIC ACID S-[Z-(a-HY- DROXY-p-TOLYL) HYDRAZIDE] (AGARITINE) A. p-Carbomethoxyphenylhydrazine A solution of 20.0 g. of p-carboxyphenylhydrazine in 200 ml. of absolute methanol and 20 ml. of concentrated sulfuric acid was heated under reflux for one hour. The solution was concentrated under reduced pressure to a volume of 75 m1. and 400 g. of ice was added to the residue. The resulting solution was cooled to -10 C. in an ice-salt bath while being treated with 30% aqueous sodium hydroxide solution. The product crystallized when the solution became strongly alkaline and it was promptly collected on a funnel and washed with ice-water until neutral. The dried product Weighed 14.3 g, (66% On recrystallization from ether, with charcoal treatment, the product, p-carbomethoxyphenylhydrazine, melted at C. (dec.).

Analysis.-Calcd. for C H N O N, 16.86. Found: N, 16.78.

B. a-Hydroxy-p-tolyllzydrazine A solution of 2.08 g. (0.0125 mole) of .p-carbomethoxyphenylhydrazine in ml. of ether (freshly distilled over lithium aluminum hydride) was added to a boiling suspension of lithium aluminum hydride (1.00 g.) in 25 ml. of dry ether over a period of 10 minutes. The reaction mixture was stirred under reflux for 1.5 hours, then allowed to remain at 25 C. for 30 minutes. The apparatus was flushed with nitrogen while 5 ml. of saturated sodium chloride solution was added in one portion. Inorganic salts were collected by filtration of the mixture, resuspendcd in ether with anhydrous sodium sulfate, and again collected and Washed with ether. The combined filtrates and washings were immediately cooled in an ice bath. The product in ether solution was a-hydroxy-ptolylhydrazine (p-methylolphenylhydrazine).

C. L-N-carbobenzyloxyglutamic acid 5-azide A solution of 1.77 g. (0.006 mole) of L-N-carbobenzyloxyglutamic acid 5-hydrazide in 25 ml. of 3 N HCl was cooled in an ice bath and then covered with ml. of cold (10 C.) ether. The cold mixture was stirred while being treated with a cold solution of 0.580 g. (0.008 mole) of sodium nitrite in 25 ml. of water over a period of 10 minutes. The lower layer was separated and extracted with two 100-ml. portions of cold (--10 C.) ether. The ether extracts containing L-N-carbobenzyloxyglutamic acid S-azide were washed with two 75-m1. portions of ice water, then combined and stored in an acetone-solid carbon dioxide hath.

D. L-N-carbobenzyloxyglutamic acid S-[Z-(a-hydroxyp-tlyl)hydrazide] The washed ether extracts of Part C were added to the ether solution of a-hydroxy-p-tolylhydrazine of Part B over a period of 60 minutes. The reaction mixture was stirred in an ice bath under nitrogen and then was main tained at a temperature of 25 C. for two hours. The resulting product in ether solution was L-N-carbobenzyloxyglutamic acid -[2-(a-hydroxy-p-tolyl)hydrazide].

E. L-glutamt'c acid 5-[Z-(a-hydroxy-pdolyl)hydrazide] (agaritine) The ether solution of L-N-carbolbenzyloxyglutarnic acid 5-[2 (a-hydroxy p tolyl)hydrazide] obtained in Part D was evaporated under reduced pressure at 30 C. almost to dryness. The residue was added to 40' ml. of methanol and 25 ml. of water and the resulting solution was hydrogenolyzed in the presence of 350 mg. of palladium-charcoal catalyst for one hour at psi. (gauge). The catalyst was removed by filtration and washed with 50 ml. of warm 50% methanol. The combined filtrate and washings were evaporated under reduced pressure at C. to a volume of 10 ml. The concentrate was diluted with 50 ml. of water. The resulting solution was extracted with two 30-m1. portions of ethyl acetate and then with two 30-ml. portions of ether.

A column (4 in. x 22 in.) was charged with 5 l. of Dowex 50, a sulfonated styrene copolymer cation-exchange resin prepared by sulfonation of copolymers of styrene and sold by Dow Chemical Company. The resin was converted to the ammonium phase by passage of dilute ammonium hydroxide through the column, after which the resin was washed with deionized water until the pH of the eflluent was approximately 7.0. The above aqueous solution, after extraction with ethyl acetate and ether as described, was applied to the column, which was then eluted with deionized water. After a forerun of 2.0 1., fractions of 500 ml. volume were collected and the ultraviolet absorption spectrum of each fraction was taken. Fractions 15, 16, and 17, having 7 max. 236 and 280 mu, were combined and evaporated under reduced pressure at C. to a volume of 7 ml. n-Butyl alcohol (25 ml.) was added and the mixture was evaporated under reduced pressure until a single phase resulted. This solu tion, left in the refrigerator, deposited 50 mg. (3%) of crystalline :agaritine with M.P. 205-9 C. (dec.). This sample was identical with natural agaritine: it had y max. 237 and 280 III/.L [04 7.1 (c. 1.14 in water), infrared spectrum identical to natural agaritine, and behavior on papergrams identical to natural agaritine.

Analysis.calcd for C12H17N3041 C, H, N, 15.72. Found: C, 53.92; H, 6.57; N, 15.45.

Following the procedure of Example 1, Parts A and B, but starting with 4-methyl L-aspartate instead of 5- methyl L-glutamate, there is obtained a cold ether solution of L-N-carbobenzyloxyaspartic acid 4-azide. Following the procedure of Example 3, Parts D and E, but subsittuting a cold ether solution of L-N-carbobenzyloxyaspartic acid 4-azide for a cold ether solution of L-N-carbobenzyloxyglutarnic acid 5-azide, there is obtained L-N-canbobenzyloxyaspartic acid 4-[2-(a-hydroxy-p-tolyl) hydrazide] which is converted by catalytic hydrogenolysis to L-aspartic acid 4-[2-(a-hydroxy-ptolyl) hydrazide].

10 EXAMPLE 4.L-GLUTAMIC ACID 5 Z-p-CAR- BOXYPHENYLHYDRAZIDE) In the same manner as shown in Example 1, Parts C and D, L-glutamic acid 5-(Z-p-carboxyphenylhydrazide) is prepared by using ,p-carboxyphenylhydrazine instead of phenylhydrazi-ne to produce L-N-carbobenzyloxyglutamic acid 5-(2-p-canboxyphenylhydrazide) and subjecting the latter compound to hydrogenolysis.

EXAMPLE 5.-L-GIJUTAMIC ACID 5-(2-p-CHLO- RO-PHENYLHYDRAZIDE) In the same manner as shown in Example 1, Parts C and D, L-glut'amic acid 5-(2-p-chlorophenylhydrazide) is prepared by using p-ohlorophenylhydrazine instead of phenylhydrazine to produce L-N-carbolbenzyloxyglutamic acid 5-(2-p-chlorophenylhydrazide) and subjecting the latter compound to hydrogenolysis.

Similarly, L-N-carbobenzyloxyglutamic acid 5-(2-p-bromophenylhydrazide), L-N-carbobenzyloxyglutamic acid 5-(2-o-bromopenylhydrazide), L-N-carbobenzyloxyglutamic acid 5-(2-p-iodophenylhydrazide), and L-N-carbobenzyloxyglutamic acid 5-[2-(3,5-dichlorophenyl)hydrazide] are prepared by using p-bromophenylhydrazine, obromophenylhydrazine, p-iodophenylhydrazine, and 3,5- dichlorophenylhydrazine instead of p-chlorophenylhydrazine. Subjecting L-N-carbobenzyloxyglutamic acid 5-(2 p-bromophenylhydrazide), L-N-carbobenzyloxyglutamic acid 5-(2-o-bromophenylhydrazide) L-N-carbobenzyloxyglutamic acid 5-(2-p-iodophenylhydrazide), and L-N- carbobenzyloxyglutamic acid 5-[2-(3,5-dichlorophenyl) hydrazide] to hydrogenolysis results in the production of L-glutamic acid 5-(Z-p-bromophenylhydrazide), L-glutamic acid 5-(2-o-bromophenylhydrazide), L-glutamic acid 5-(Z-p-iodophenylhydrazide), and L-glutamic acid 5- [2- (3 ,5 -dichlorophenyl hydrazide] respectively.

EXAMPLE 6.AMINOMALONIC ACID 2-0- METHOXYPHENYLHYDRAZIDE A-. N-carbobenzyloxyaminomalonic acid 2-a-methoxyphenylhydrazide In the same manner as shown in Example 1, Parts A, B and C, N-carbobenzyloxyaminomalonic acid 2-o-methoxyphenylhydrazide is prepared by using methyl aminomalo nate and o-methoxyphenylhydrazine instead of S-methyl L-glutarnate and phenylhydrazine, respectively.

B. Aminomalom'c acid 2-o-methoxyphenylhydrabide In the same manner as shown in Example 1, Part D, aminomalonic acid 2-o-methoxyphenylhydrazide is prepared by using N-carbobenzyloxyaminomalonic acid 2-0- methoxyphenylhydrazide instead of L-N-carbobenzyloxyglutamic acid 5-(2-phenylhydrazide).

Similarly, N-carbobenzyloxyarninomalonic acid 2-0- chlorophenylhydrazide, aminomalonic acid 2-o-chlorophenylhydrazide; N-carbobenzyloxyaminomalonic acid 2- o-tolylhydrazide, aminomalonic acid 2-o-tolylhydrazide; N-carbobenzyloxyaminomalonic acid 2-p-propoxyphenylhydrazide, aminomalonic acid Z-p-propoxyphenylhydrazide; N-carbobenzyloxyaminomalonic acid 2-p-iodophenylhydrazide and aminomalonic acid Z-p-iodo-phenylhydrazide are prepared by utilizing o-chlorophenylhydrazine, o-tolylhydrazine, p-propoxyphenylhydrazine, and p-iodophenylhydrazine instead of o-methoxyphenylhydrazine.

Substituting allyl chloroformate and p-toluenesulfonyl chloride for benzyl chloroformate results in the production of N-carboallyloxyaminomalonic acid 2-o-methoxyphenylhydrazide, N-tosylaminomalonic acid 2-o-methoxyphenylhydrazide; N-carboallyloxyaminomalonic acid 2- o-chlorophenylhydrazide, N-tosylaminomalonic acid 2- o-chlorophenylhydrazide; N-carboallyloxyaminomalonic acid 2-o-tolylhydrazide, N-tosylaminomalonic acid 2-otolylhydrazide; N-carboallyloxyaminomalonic acid 2-ppropoxyphenylhydrazide, N-tosylaminomalonic acid 2-p- 11 propoxyphenylhydrazide; N-carboallyloxyaminomalonic acid 2-p-iodophenylhydrazide, and N-tosylaminomalonic acid 2-p-iodophenylhydrazide.

EXAMPLE 7.L-ASPARTIC ACID 4-(2-p- ETHOXYPHENYLHYDRAZIDE) A. L-N-carboallyloxyaspartic acid 4-(2-p-eth0xyphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, L-N-carboallyloxyaspartic acid 4-(2-p-ethoxyphenylhydrazide) is prepared by using allyl chloroformate, 4-rnethyl L-aspartate, and p-ethoxyphenylhydrazine instead of benzyl chloroformate, S-methyl L-glutamate and phenylhydrazine, respectively.

B. L-aspartic acid 4-(Z-p-ethoxyphenylhydrazide) EXAMPLE 8.3,3-DIMETHYLASPARTIC ACID 4- (Z-m-PROPOXYPHENYLHYDRAZIDE) A. N-tsyl-3,3-dimethylaspartic acid 4-(2-m-pr0poxyphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-tosyl-3,3-dimethylaspartic acid 4-(2-m-propoxyphenylhydrazide) is prepared by using p-toluenesulfonyl chloride, 4-methyl 3,3-dimethylaspartate, and mpropoxyphenylhydrazine instead of benzyl chloroformate, S-methyl L-glu-tamate and phenylhydrazine, respectively.

B. 3,3-dimethylaspartic acid 4-(2-m-pr0p0xyphenylhydrazide) N-tosyl 3,3-dimethyl-aspartic acid 4-(2-m-propoxyphenylhydrazide) is reacted with sodium and liquid ammonia in accordance with the procedure described in J. Amer. Chem. Soc. 79, 639-44, 1957, to produce 3,3-dimethyl-aspartic acid 4-(2-m-propoxyphenylhydrazide).

Similarly, N-tosyl-3,3-dimethylaspartic acid 4-(2-piodophenylhydrazide) and 3,3-dimethylaspartic acid 4-(2- p-iodophenylhydrazide) are prepared by utilizing p-iodophenylhydrazine instead of m-propoxyphenylhydrazine.

EXAMPLE 9.2-PROPYLASPARTIC ACID 4- [2- (3,4, S-TRIMETI-IOXYPHENYL HYDRAZIDE] A-. N-carbobenzyl0xy-2-pr0pylaspartic acid 4- [2-(3,4,5- trimethoxyphenyl hydrazide] In the same manner as shown in Example 1, Parts A, B and C, N-carbobenzyloxy-Z-propylaspartic acid 4-[2-(3, 4,5-trimethoxyphenyl)hydrazide] is prepared by using 4- methyl 2-propylaspartate and 3,4,5-trimethoxyphenylhydrazine instead of S-methyl L-glutamate and phenylhydrazine, respectively.

B. Z-propylaspartic acid 4-[2-(3,4,5-trimethoxyphenyl) hydrazide] In the same manner as shown in Example 1, Part D, 2 propylaspartic acid 4 [2 (3,4,5 trimethoxyphenyl) hydrazide] is prepared by using N carbobenzyloxy 2- propylaspartic acid 4 [2 (3,4,5 trimethoxyphenyl)hydrazide] instead of N- carbobenzyloxyglutamic acid 5- (2-phenylhydrazide) I .2. EXAMPLE 10.-'3 -BUTYLASPARTIC ACID 4- 2-p- ETHYLPHENYLHYDRAZIDE) A. N-carbobenzyloxy-3-butylaspartic acid 4-(2-pethylphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-carbobenzyloxy-3-butylaspartic acid 4- (Z-p-ethylphenylhydrazide) is prepared by using 4-methyl 3-butylaspartate and p-ethylphenylhydrazine instead of S methyI L-glutamate and phenylhy-drazine, respectively.

B. 3-butylaspartic acid 4-(Z-p-eihylphenylhydnazide) In the same manner as shown in Example 1, Part D, 3-butylaspartic acid 4-(2-p-ethylphenylhydrazide) is prepared by using N-carbobenzyloxy-3-butylaspartic acid 4 (2 p ethylphenylhydrazide) instead of L N carbobenzyloxyglutamic acid 5-(2-phenylhydrazide).

EXAMPLE 1l.--3-METHYLASPARTIC ACID 4- (2-p-PROPYLPHENYLHYDRAZIDE) A. N-t0syl-3- mezhylaspartic acid 4-(2-ppropylphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-tosyl-3-methy1aspartic acid 4-(2-p-propylphenylhydrazide) is prepared by using p-toluenesulfonyl chloride, 4-methyl 3-methylaspartate, and p-propylphenylhydrazine instead of benzyl chloroformate, S-methyl L- glutamate, and phenylhydrazine, respectively.

B. 3-methylaspartic acid 4-(2-p-pr0pylphenylhydrazide) N-tosyl-3-methylaspartic acid 4-(2-p-propylphenylhydrazide) is reacted with sodium and liquid ammonia in accordance with the procedure described in J. Amer. Chem. Soc. 79, 63944, 1957, to produce 3-methylaspartic acid 4-(2-p-propylphenylhydrazidc).

EXAMPLE 12.2-METHYLGL'UTAMIC ACID 5- (2-p-BUTYLPHENYLI-IYDRAZIDE) A. N-carboallyloxy-Z-methylglutamic acid 5-(2-pbutylpkenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-carboallyloxy-2-methylglutamic acid 5- (2-p+buty1phenylhydrazide) is prepared by using allyl chloroformate, S-methyl Z-methylglutamate, and p-butylphenylhydrazine instead of benzyl chloroformate, 5- methyl L-glutamate, and phenylhydrazine, respectively.

B. Z-methylglutamic acid 5-(2-p-l atylphenylhydnazide) N carboallyloxy 2 methylglutamic acid 5 (2 pbutylphenylhydrazide) is hydrogenolyzed in the presence of platinum oxide catalyst in acidified ethanol in accordance With the procedure described in J. Amer. Chem. Soc. 79, 63739, 1957, to produce Z-methylglutamic acid 5- 2-butylphenylhydrazide) EXAMPLE l3 .3 -METHYLGLUTAMIC ACID 5- (2-0-ISOPROPYLPHENYLHYDRAZIDE) A. N-carb0allyl0xy-3-mezhylglutamic acid 5-(2-0- isop ropylphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-carboallyloXy-3-methylglutamic acid 5- (2-0-isopropylphenylhydrazide) is prepared by using allyl chloroformate, S-methyl 3-methylglutamate, and o-isopropylphenylhydrazine instead of benzyl chloroformate, S-methyl L-glutamate, and phenylhydrazine, respectively.

B. 3-i1iethylglutamic acid 5-(2-0- isopropylphenylhydrazide) N carboallyloxy 3 methylglutamic acid 5 (2 oisopropylphenylhydrazide) is hydrogenolyzed in the presence of platinum oxide catalyst in acidified ethanol in accordance with the procedure described in J. Amer. Chem. Soc. 79, 637-39, 1957, to produce 3-methylg1utamic acid 5-(2-o-isopropylphenylhydrazide).

13 EXAMPLE 14.4-METHYLGLUTAMIC ACID 5 (Z-PHENYLHYD RAZIDE) A. N-tsyl-4-methylglulamic acid S-(Z-phenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-tosyl-4-methylglutamic acid S-(Z-phenylhydrazide) is prepared by using p-toluenesulfonyl chloride and S-methyl 4-methylglutamate instead of benzyl chloroformate and S-methyl L-glutamate, respectively.

B. 4-methylglutamic acid -(2-phenylhydrwzide) N-tosyl-4-methylglutamic acid 5-(2-phenylhydrazide) is reacted with sodium and liquid ammonia in accordance with the procedure described in J. Amer. Chem. Soc. 79, 639-44, 1957, to produce 4-methylglutamic acid 5- (2-phenylhydrazide) EXAMPLE .2-AMINOHEXANEDIOIC ACID 6- (Z-P-CHLOROPHENYLHYD RAZIDE) A. N-carboallyloxy-Z-aminohexanedioic acid 6-(2-pchlorophenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-carboallyloxy-Z-aminohexanedioic acid 6-(2-p-chlorophenylhydrazide) is prepared by using allyl chloroformate, 6-methyl Z-aminohexanedioate, and pchlorophenylhydrazine instead of benzyl chloroformate, S-methyl L-glutamate and phenylhydrazine, respectively.

B. Z-aminohexanedioz'c acid 6-(2-pchlorophenylhydrazide) N-carboallyloxy-Z-aminohexanedioic acid 6-(2-p-chlorophenylhydrazide) is hydrogenolyzed in the presence of platinum oxide catalyst in acidified ethanol in accordance with the procedure described in J. Amer. Chem. Soc. 79, 637-39, 1957, to produce 2-aminohexanedioic acid 6- 2-p-chlorophenylhydrazide) Similarly,

N-carboallyloxy-2-arninohexanedioic acid 6-(2-0- ethylphenylhydrazide),

2-aminohexanedioic acid 6-(2-o-ethylphenylhydrazide);

N-carboallyloxy-2-aminohexanedioic acid 6-(2-mpropoxyphenylhydrazide) Z-aminohexanedioic acid 6-(2-m-propoxyphenylhydrazide);

N-carboallyloxy-2-aminohexanedioic acid 6-(2-mfluorophenylhydrazide) and 2-aminohexanedioic acid 6-(2-m-fluorophenylhydrazide) are prepared by substituting o-ethylphenylhydrazine, mpropoxyphenylhydrazine, and m-fluorophenylhydrazine for p-chlorophenylhydrazine.

Substituting benzyl chloroformate and p-toluenesulfonyl chloride for allyl chloroformate results in the production of 14 EXAMPLE 16.2-AMINOHEPTANEDIOIC ACID 7-(2-p-METHYLOLPHENYLHYDRAZIDE) A. N -carb0allyloxy-Z-amin0heptanedi0ic acid 7-(2-p-methyl0lphenylhydrazide) Following the procedure of Example 1, Parts A and B, but substituting allyl chloroformate and 7-methyl 2- aminoheptanedioate for benzyl chloroformate and S-methyl L-glutamate, respectively, and using the resulting cold ether solution of N-carboallyloxy-Z-aminoheptanedioic acid 7-azide in the procedure of Example 3, Part D, N- carboallyloxy-2-aminoheptanedioic acid 7-(2-p-methylolphenylhydrazide) is obtained.

B. Z-aminoheptanedioic acid 7-(2-pmethylolphenylhydrazide) N-carboallyloxy-Z-aminoheptanedioic acid 7-(2-p-methylolphenylhydrazide) is hydrogenolyzed in the presence of platinum oxide catalyst in acidified ethanol in accordance with the procedure described in J. Amer. Chem. Soc. 79, 637-39, 1957, to produce Z-aminoheptanedioic acid 7-(2- p-methylolphenylhydrazide) Similarly,

N-carb0allyloxy-2-aminoheptanedioic acid 7-(2-ptolylhydrazide 2-aminoheptanedioic acid 7-(2-p-tolylhydrazide);

N-carboa11yloxy-2-aminoheptanedioic acid 7 -(2-ppropoxyphenylhydrazide) Z-aminoheptanedioic acid 7-(2-p-propoxyphenylhydrazide);

N-carboallyloxy-Z-aminoheptanedioic acid 7-(2-pchlorophenylhydrazide) and Z-aminoheptanedioic :acid 7-(2 p-chlorophenylhydrazide) are prepared by utilizing p-tolylhydrazine, p-propoxyphenylhydrazine and p-chlorophenylhydrazine instead of p-methylolphenylhydrazine.

Substituting benzyl chloroformate and p-toluenesulfonyl chloride for allyl chloroformate results in the production of N-carbobenzyloxy-Z-aminoheptanedioic acid 7-(2-pmethylolphenylhydrazide N-tosyl-2-aminoheptanedioic acid 7-(2-p-methylolphenylhydrazide) N-carbobenzyloxy-Z-aminoheptanedioic acid 7-(2-ptolylhydrazide) N-tosyI-Z-aminoheptanedioic acid 7-(2-p-tolylhydrazide); N-carbobenzyloxy-2-aminoheptanedioic acid 7-(2-ppropoxyphenylhydrazide N-tosyl-Z-aminoheptanedioic acid 7-(2-p-propoxyphenylhydrazide) N-carbobenzyloxy-Z-aminoheptanedioic acid 7- (2-pchlorophenylhydrazide) and N-tosyl-Z-aminoheptanedioic acid 7-(2-p-chlorophenylhydrazide) EXAMPLE l7.-2-AMINOOCTANEDIOIC ACID 8-(Z-m-CARBOXYPHENYLHYDRAZIDE) A. N-carbobenzyloxy-Z-amin00ctanedi0ic acid 7 8- (Z-m-carboxyphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-carbobenzyloxy-2-aminooctanedioic acid 8- (2-m-carboxyphenylhydrazide) is prepared by using 8 methyl 2-aminooctanedioate and m-carboxyphenylhydrazine instead of S-methyl L-glutamate and phenylhydrazine, respectively.

B. Z-aminooctanedioic acid 8-(2-mcarboxyphenylhydrazide) In the same manner as shown in Example 1, Part D, 2-aminooctanedioic acid 8-(2-m-carboxyphenylhydrazide) is prepared by using N-carbobenzyloxy-Z-aminooctanedioic acid 8-(Z-m-carboxyphenylhydrazide) instead of L N-carbobenzyl-oxyglutamic acid 5-(2-phenylhydrazide).

Similarly,

N-carbobenzyloxy-Z-aminooctanedioic acid 8-(2-pethylphenylhydrazide) Z-aminooctanedioic acid 8-(Z-p-ethylphenylhydrazide);

N-carbobenzyloxy-Z-aminooctanedioic acid 8-(2-mbutoxyphenylhydrazide) Z-aminooctanedioic acid 8-(2-m-butoxyphenylhydrazide);

N-carbobenzyloxy-Z-aminooctanedioic acid 8-(2-0- chlorophenylhydrazide) 2-aminooctanedioic acid 8-(2-o-chlorophenylhydrazide) are prepared by utilizing p-ethylphenylhydrazine, mbutoxyphenylhydrazine, and o-chlorophenylhydrazine instead of m-carboxyphenylhydrazine.

Substituting allyl chloroformate and p-toluenesulfonyl chloride for benzyl chloroformate results in the production of N-carboallyloxy-Z-aminooctanedioic acid 8-(2-mcarboxyphenylhydrazide I N-tosyl-Z-aminooctanedioic acid 8-(2-m-carboxyphenylhydrazide) N-carboallyloxy-Z-aminooctanedioic acid 8-(2-pethylphenylhydrazide) N-tosyl-2-aminooctanedioic acid 8-(2-p-ethylphenylhydrazide); N-carboallyloxy-2-aminooctanedioic acid 8-(2-mbutoxyphenylhydrazide) N-tosyl-Z-aminooctanedioic acid 8-(2-mbutoxyphenylhydrazide) N-carboallyloxy-Z-aminooctanedioic acid 8-(2-0- chlorophenylhydrazide) and N-tosyl-Z-aminooctanedioic acid 8-(2-0- chlorophenylhydrazide) EXAMPLE 1 8.AMINONONANEDIOIC AOI'D 9 (2-m-IOD O PHENYLHYDRAZIDE) A. N-tosyl-Z-aminn0nanedioic acid 9-(2-miodophenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-tosyl-2-aminononanedioic acid 9-(2-m-i0dophenylhydrazide) is prepared by using p-toluenesulfonyl chloride, 9-methyl Z-aminononanedioate, and m-iodophenylhydrazine instead of benzyl chloroformate, S-methyl L- glutamate, and phenylhydrazine, respectively.

B. Z-aminononanedioic acid 9-(2-m-i0d0phenylhydrazide) N-tosyl-2-aminononanedioic acid 9-(2-m-iodophenylhydrazide) is reacted with sodium and liquid ammonia in accordance with the procedure described in J. Amer. Chem. Soc. 79, 63944, 1957, to produce 2-aminononanedioic acid 9-(2-m-iodophenylhydrazide).

Similarly, N-tosyl-2-aminononanedioic acid 9-(2-ppropylphenylhydrazide), Z-aminononanedioic acid 9-(2-ppropylphenylhydrazide); N-to'syl-2-aminononanedioic acid 9-(2-o-butoxyphenylhydrazide), Z-aminononanedioic acid 9- (2-o-butoxyphenylhydrazide) N-tosyl-Z-aminononanedioic acid 9-(2-p-chlorophenylhydrazide) and 2-aminononanedioic acid 9-(2-p-chlorophenylhydrazide) are prepared by utilizing p-propylphenylhydrazine, o-butoxyphenylhydrazine, and p-chlorophenylhydrazine instead of miodophenylhydrazine.

Substituting benzyl chloroformate and allyl chloroformate for p-toluenesulfonyl chloride results in the production of N-carbobenzyloxy-Z-aminononanedioic acid 9- (2 rn iodophenylhydrazide), N-carboallyloxy-Z-aminononanedioic acid 9-(2-m-iodophenylhydrazide); N-carbobenzyloxy-Z-aminononanedioic acid 9-(2-p-propylphenylhydrazide), N-carboallyloxy-Z-aminononanedioic acid 9- (2-p-propylphenylhydrazide); N-carbobenzyloxy-Z-aminononanedioic acid 9-(2-o-butoxyphenylhydrazide), N-carboallyloxy-Z-aminononanedioic acid 9-(2-o-butoxyphenylhydrazide) N-carbobenzyloxy-Z-aminononanedioic acid 16 9 (2-p-chlorophenylhydrazide) and N-carboallyloxy-Z- aminononanedioic acid 9- (2-p-chloropheny1hydrazide) EXAMPLE 19.2-AMINODECANEDIOIC ACID 10- (Z- -METHOXYPHENYLHYDRAZIDE) A. N-tosyZ-Z-aminodecanedioic acid 10-(2-pmethoxyphenylhydrazide) In the same manner as shown in Example 1, Parts A, B and C, N-tosyl-2-aminodecanedioic acid 10(2-p-methoxyphenyl-hydrazide) is prepared by using p-toluenesulfonyl chloride, IO-methyl 2-aminodecanedioate, and prnethoxyphenylhydrazine instead of benzyl chloroformate, S-methyl L-glutamate, and phenylhydrazine, respectively.

B. Z-aminodecanedioic acid IO-(Z-p-methoxyphenylhydrazide) N-tosyl-2-aminodecanedioic acid 10-(2-p-methoxyphenylhydrazide) is reacted wtih sodium and liquid ammonia in accordance with the procedure described in J. Amer. Chem. Soc. 79, 639-44, 1957, to produce Z-aminodecanedioic acid 10-(2-p-methoxyphenylhydrazide).

Similarly, N-tosyl-2-aminodecanedioic acid 10-(2-ppropylphenylhydrazide), Z-aminodecanedioic acid l0-(2-p propylphenylhydrazide); N-tosyl-2-aminodecanedioic acid 10 (2-p-carboxyphenylhydrazide), 2-aminodecanedioic acid 10-(Z-p-carboxyphenylhydrazide); N-tosyl-2-amino- "decanedioic acid 10-(2-p-iodophenylhydrazide) and 2- aminodecanedioic acid 10-(2-p-iodophenylhydrazide) are prepared by utilizing p-propylphenylhydrazine, p-carboxyp-henylhydrazine, and p-iodophenylhydrazine instead of pmethoxyphenylhydrazine.

Substituting benzyl chloroformate and allyl chloroformate for p-toluenesulfonyl chloride results in the production of N-carbobenzyloxy-2-aminodecanedioic acid 10- (2-p-methoxyphenylhydrazide) N-carboallyloXy-2-aminodecanedioic acid 10-(2-p-methoxyphenylhydrazide); N- car-bobenzyloxy-Z-aminodecanedioic acid 10-(2-p-propylphenylhydrazide), N carboallyloxy-Z-aminodecanedicic acid 10-(2-p-propylphenylhydrazide); N-carbobenzyloxy- 2 aminodecanedioic acid 10-(2-p-carboxyphenylhydrazide), N-carboallyloxy-Z-aminodecanedioic acid 10-(2-pcarboxyphenylhydrazide); N carbobenzyloxy-Z-aminodecanedioic acid 10-(2-p-iodophenylhydrazide) and N carboallyloXy-2-aminodecanedioic acid 10-(2-p-iodophenylhydrazide) We claim:

1. Compounds represented by the formula:

wherein R is selected from the group consisting of hydrogen, methylol, halogen, carboxy, alkyl of 1 to 4 carbon atoms, inclusive, and alkoxy of 1 to 4 carbon atoms, inclusive, R is selected from the group consisting of hydrogen and alkyl of 1 to 4 carbon atoms, inclusive, R is selected from the group consisting of carboallyloxy, carbobenzyloxy and tosyl, and n is an integer from zero to 7, inclusive.

dZ.) L-N-carbobenzyloxyglutamic acid 5-(2-phenylhydra- Z1 e 3. L-N-carbobenzyloxyglutamic acid 5-[2-(a-hydroxyptolyl)hydrazide].

4. L N carbobenzyloxyglutamic acid 5-(2-p-toly1hydrazide).

References Cited by the Examiner Hofmann et a1.: J. Amer. Chem. Soc. 74, pp. 470-72 (1952).

Leven'berg: J. Amer. Chem. Soc. 83, p. 503 (1961).

LORRAINE A. WEINBERGER, Primaiy Examiner.

L. A. THAXTON, Assistant Examiner. 

1. COMPOUNDS REPRESENTED BY THE FORMULA: (R3-PHENYL)-NH-NH-CO-(CNH2N)-C(-R'')(-NH-R2)-COOH WHEREIN R3 IS SELECTED FROM THE GROUP CONSISTING OF HYDROGEN, METHYLOL, HALOGEN, CARBOXY, ALKYL OF 1 TO 4 CARBON ATOMS, INCLUSIVE, AND ALKOXY OF 1 TO 4 CARBON ATOMS, INCLUSIVE, R1 IS SELECTED FROM THE GROUP CONSISTING OF HYDROGEN AND ALKYL OF 1 TO 4 CARBON ATOMS, INCLUSIVE, R2 IS SELECTED FROM THE GROUP CONSISTING OF CARBOALLYLOXY, CARBOBENZYLOXY AND TOSYL, AND N IS AN INTEGER FROM ZERO TO 7, INCLUSIVE. 